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1. Salmonella Bacterin Against Probiotic
Salmonella Enteritidis Infection in Broiler

2. Trial Aim
This study was carried out to investigate the efficacy of the
locally prepared autogenous Salmonella Enteritidis (S.
Enteritidis) bacterin as well as a probiotic preparation in the
prevention of broiler chickens from S. Enteritidis infection.

3. Experimental Chickens
310, day-old Hubbard
broiler chicks of mixed sex.
Obtained from Cairo
Poultry Company in 10th
Ramadan city.
Breeder flock free from
salmonellosis.

4. Experimental Design
 At arrival, randomly 10 chicks were sacrificed and then
examined bacteriologically to prove their freedom from S.
Enteritidis infection.

5. Experimental Chickens
The birds were kept under complete observation for six weeks
experimental period) in separate thoroughly cleaned and
disinfected houses.

6. Vaccination Program
ND IB AI IBD
Day 6: HB1
Day 19: L aSota
Day 6: H120 Day 7: H5N1 RG Day 14: 228E

7. Salmonella Vaccination Program
 Inactivated S. Enteritidis bacterin was given IM in the thigh
muscle for the experimental chicks at:
1. First day of age in a dose of 0.2 ml/bird
2. Second dose at 10 days of age in a dose 0.5 ml/bird.

8. The Used Probiotic
 Commercial preparation containing:
– (Lactobacillus acidophilus, Enterococcus faecium, Lactobacillus
plantarum and Lactobacillus casei)
– Potassium
– Vitamins A, D3, E and K,
– Riboflavin, pantothenic acid, thiamine and niacinamide.
 That product was manufactured by Bomac Vets Plus, USA.
Batch No., A704.
 It was given in the drinking water at the age of one day for 5
consecutive days in a dose of 1gm/ 4 liter of the drinking
water as recommended by manufacturer.

9. The Challenge Inoculum
 Broth culture of S. Enteritidis field strain was centrifuged at
3000 r.p.m for 10 min.
 Sediment was diluted with sterile buffer saline and adjusted
using Mac Ferland matching tube to contain 10 log 9 CFU/ml.
 The challenge inoculum was prepared according to the
method of Timms et al., (1990).
 At 20 days of age, each bird in the experimentally infected
groups was inoculated orally with 0.5 ml/ containing 10 log 9
CFU/ml S. Enteritidis (Okamoto et al., 2007).

10. 4 Groups, each of 75 chicks:
Non-Vaccinated
Non-Challenged
1
Infected
Not Treated
2
Vaccinated
Infected
3
Probiotic
Infected
4

11. Evaluation
1. Clinical Signs
2. Mortalities
3. Performance
4. Gross Lesions
5. Detection of the Shedding of S. Enteritidis
Birds in the challenged groups were observed daily for three
weeks post challenge till the end of the study (6 weeks of age)
for the clinical signs or deaths.
Dead birds were subjected to necropsy for recording the lesions
of S. Enteritidis (O'Brien 1988).

12. Cont. …
Detection of the Shedding of S. Enteritidis
 Cloacal swabs were taken from birds in each group just before
experimental infection (at 20 days of age) to ensure that the birds free
from S. Enteritidis infection.
 Weekly after the challenge up to 6 weeks of age, cloacal swabs were
collected from each of the infected as well as control group and examined
bacteriologically for the presence of S. Enteritidis organism.
 Sterile cotton swab was inserted into the cloaca of each bird and rotated
gently to collect the clocal contents.
 Each swab was transferred to 10 ml tube of tetrathionate broth and
incubated overnight at 37°C.
 A loopful from the broth was streaked on S.S agar for Salmonella isolation.
 Suspected colonies were identified morphologically and biochemically.

13. Cont. …
Re-isolation of S. Enteritidis
 Ten birds from each group post challenge were weekly
randomly selected, sacrificed and the liver, heart, spleen and
caecum were collected for S. Enteritidis re-isolation.
 Samples were inoculated into tetrathionate broth, incubated
at 37°C for 24 hr, streaked onto S.S agar and incubated at 37°C
for 24 hr. Suspected colonies were identified morphologically
and biochemically.

14. Cont. …
Performance
At arrival, the chicks were weighed and then the birds in each
group were subjected to weekly determination of the production
parameters that include;
1. Body weight (BW)
2. Cumulative feed conversion (CFC)
3. European production efficiency factor (EPEF) according to
sainsbury (1984).
These measures were taken till the end of the study (6 weeks of
age).

15. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0
23
4
9
Number of Dead Birds

16. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
31%
5%
12%
Mortality Rate %

17. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
52%
18%
26%
SE Faecal Shedding Week 1

18. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
37%
7%
18%
SE Faecal Shedding Week 2

19. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
25%
0%
4%
SE Faecal Shedding Week 3

20. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
41%
9%
18%
SE Faecal Shedding Total
SE Faecal Shedding Total

21. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
0%
47%
12%
22%
Re-isolation rate of S. Enteritidis from different organs

22. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
2.03
2.43
1.78
1.89
Cumulative Feed Conversion

23. Results
Control Non-treated -
Infected
Vaccinated -
Infected
Probiotic -
Infected
1,580
1,355
1,705
1,640
Average body weight after 6 weeks

24. Conclusion
From this study, it could be concluded that both the locally
prepared autogenous S. Enteritidis bacterin in double doses and
the probiotic preparation are effective and safe methods for
prevention of S. Enteritidis infection in broiler chickens.